Production and biochemical and biophysical characterization of fibrinolytic protease of a Mucorsubtilissimus strain isolated from the caatinga biome

被引：0

作者：

Conniff, Amanda Emmanuelle S. ^{[1
,2
]}

Nascimento, Thiago P. ^{[3
]}

Costa, Romero Marcos P. B. ^{[4
]}

Breydo, Leonid ^{[5
]}

Porto, Camila S. ^{[6
]}

Converti, Attilio ^{[7
]}

Siqueira, Joyce G. W. ^{[8
]}

Teixeira, Jose Antonio ^{[9
]}

de Campos-Takaki, Galba Maria ^{[10
]}

Uversky, Vladimir N. ^{[5
]}

Porto, Ana Lucia F. ^{[1
]}

Porto, Tatiana S. ^{[1
]}

机构：

[1] Univ Fed Rural Pernambuco, Dept Morfol & Fisiol Anim, Rua Dom Manoel de Medeiros S-N, BR-52171900 Recife, PE, Brazil

[2] Univ S Florida, Morsani Coll Med, Dept Med Engn, 12901 Bruce B Downs Blvd, Tampa, FL 33612 USA

[3] Univ Fed Piaui, Campus Prof Cinobelina Elvas,BR 135,Km 3, BR-64049550 Bom Jesus, PI, Brazil

[4] Univ Pernambuco, Inst Ciencias Biol, Rua Arnobio Marques 310, BR-50100130 Recife, PE, Brazil

[5] Univ S Florida, Morsani Coll Med, Dept Mol Med, 12901 Bruce B Downs Blvd, Tampa, FL 33612 USA

[6] Univ Fed Alagoas, Unidade Educ Penedo, Ave Beira Rio S-N, BR-57200000 Penedo, Brazil

[7] Genoa Univ, Dept Civil Chem & Environm Engn, Pole Chem Engn, Via Opera Pia 15, I-16145 Genoa, Italy

[8] Univ Fed Rural Pernambuco, Posgrad Ciencia & Tecnol Alimentos, Rua Dom Manoel de Medeiros S-N, BR-52171900 Recife, PE, Brazil

[9] Univ Biol Minho, Inst Biotechnol & Bioengn, Ctr Engn, Campus Gualtar, Campus Gualtar, P-4710057 Braga, Portugal

[10] Univ Catolica Pernambuco, Nucleo Pesquisas Ciencias Ambientais, Rua Principe 526, BR-50050900 Recife, PE, Brazil

来源：

ANAIS DA ACADEMIA BRASILEIRA DE CIENCIAS | 2024年 / 96卷

关键词：

circular dichroism; fibrinolysis; fibrinolytic enzyme; Mucor; protease; submerged fermentation; SERINE-PROTEASE; THERMAL-DENATURATION; CIRCULAR-DICHROISM; PURIFICATION; ENZYME; FUNGUS; ANTICOAGULANT; PH;

D O I：

10.1590/0001-3765202420230616

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Cardiovascular diseases, resulting from the deposition of clots in blood vessels, are the leading cause of death worldwide. Fibrinolytic enzymatic activity can catalyze blood clot degradation. Findings show that 36 fungal isolates recovered from Caatinga soils have the potential to produce fibrinolytic protease under submerged conditions. About 58 % of the isolates displayed fibrinolytic activity above 100 U/mL, with Mucor subtilissimus UCP 1262 being the most active. The protease was biochemically and biophysically characterized, showing that the enzyme had a high affinity for SAApNA substrate and was significantly inhibited by fluoride methyl phenyl sulfonyl-C7H7FO2S, suggesting that it is a chymotrypsin-like serine protease. The highest enzyme activity was detected at pH 5.0 and 28 degrees C. This fibrinolytic protease's far-UV circular dichroism (CD) showed that its secondary structure was primarily alpha-helical. The purified fibrinolytic enzyme may represent a novel therapeutic agent for treating thrombosis. At temperatures above 65 degrees C, the enzyme lost all its secondary structure. Its melting temperature was 58.1 degrees C, the denaturation enthalpy 85.1 kcal/mol, and the denaturation entropy 0.26 kcal/K center dot mol.

引用

页数：15